生物合成
生物化学
酶
大肠杆菌
辅因子
化学
产量(工程)
黄色粘球菌
基因
突变体
冶金
材料科学
作者
Zhongkui Li,Zhijian Ni,Xiang-Song Chen,Gang Wang,Jinyong Wu,Jianming Yao
出处
期刊:Molecules
[MDPI AG]
日期:2020-08-06
卷期号:25 (16): 3567-3567
被引量:7
标识
DOI:10.3390/molecules25163567
摘要
Among the human milk oligosaccharides (HMOs), one of the most abundant oligosaccharides and has great benefits for human health is 3′-sialyllactose (3′-SL). Given its important physiological functions and the lack of cost-effective production processes, we constructed an in vitro multi-enzymatic cofactor recycling system for the biosynthesis of 3′-SL from a low-cost substrate. First, we constructed the biosynthetic pathway and increased the solubility of cytidine monophosphate kinase (CMK) with chaperones. We subsequently identified that β-galactosidase (lacZ) affects the yield of 3′-SL, and hence with the lacZ gene knocked out, a 3.3-fold increase in the production of 3′-SL was observed. Further, temperature, pH, polyphosphate concentration, and concentration of divalent metal ions for 3′-SL production were optimized. Finally, an efficient biotransformation system was established under the optimized conditions. The maximum production of 3′-SL reached 38.7 mM, and a molar yield of 97.1% from N-acetylneuraminic acid (NeuAc, sialic acid, SA) was obtained. The results demonstrate that the multi-enzymatic cascade biosynthetic pathway with cofactor regeneration holds promise as an industrial strategy for producing 3′-SL.
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